The NucleoSpin FFPE DNA kit provides a convenient, reliable, and fast method to isolate DNA from formalin-fixed, paraffin-embedded (FFPE) tissue specimens. The procedure omits the use of flammable and malodorous xylene or d-limonene commonly used for deparaffinization. Further, the procedure omits the difficult removal of organic solvent from often barely visible tissue pellets. NucleoSpin FFPE DNA employs the odorless Paraffin Dissolver (patent pending) and allows efficient lysis in a convenient two-phase system.**

First, the paraffin of FFPE sections is dissolved in the Paraffin Dissolver. Tissue is then digested by proteinase to solubilize the fixed tissue and release DNA into solution. Subsequently, heat incubation with a specially designed buffer effectively eliminates crosslinks from the previously released DNA. After addition of ethanol, the lysate is applied to the NucleoSpin FFPE DNA Column. DNA is bound to the silica membrane. Two washing steps help to remove salts, metabolites, and macromolecular cellular components. Pure DNA is finally eluted under low ionic strength conditions in a small volume (5–30 µl) of Elution Buffer BE, yielding highly concentrated DNA.

Isolated DNA is high in concentration, formaldehyde crosslinks are reversed efficiently, and the DNA quality of the sample is well preserved during the complete process (deparaffinization, decrosslink, purification). Thus, isolated DNA shows good performance in typical downstream applications like PCR.

**Users preferring xylene deparaffinization will find protocols utilizing xylene for deparaffinization in the manual. Deparaffinization using xylene shows the same efficiency as deparaffinization using Paraffin Dissolver. Xylene is not included in the kit.

Features

Overcome formalin crosslinking of DNA by incubation in decrosslinking buffer

High-quality DNA provides improved performance in PCR

Applications

Rapid isolation of DNA from formalin-fixed, paraffin-embedded samples

Isolation of DNA from fresh and archived FFPE samples

Isolation of DNA from specimen on object slides

Typical downstream application: PCR

Product Overview

Technology

Silica membrane

Format

Mini spin columns – XS design

Starting Material

<7 sections (10 µm) of 250 mm2 total area<15 mg paraffin*

Typical Yield

Depends on amount and quality of the sample

A260/280

1.90–2.10

DNA Quality

Strongly depends on sample quality

Elution Volume

5–30 µl

Preparation Time

~70 min/6 preps

Binding Capacity

110 µg

*When using the standard protocol with Paraffin Disolver. Larger quantities of paraffin can be processed when using additional Paraffin Dissolver or the standard protocol with xylene for deparaffinization.